J Periodontal Implant Sci.  2010 Jun;40(3):119-124. 10.5051/jpis.2010.40.3.119.

Leptin potentiates Prevotella intermedia lipopolysaccharide-induced production of TNF-alpha in monocyte-derived macrophages

• Kim SJ

• KMID: 2212138
• DOI: http://doi.org/10.5051/jpis.2010.40.3.119

Abstract

PURPOSE
In addition to regulating body weight, leptin is also recognized for its role in the regulation of immune function and inflammation. The purpose of this study was to investigate the effect of leptin on Prevotella (P.) intermedia lipopolysaccharide (LPS)-induced tumor necrosis factor (TNF)-alpha production in differentiated THP-1 cells, a human monocytic cell line.
METHODS
LPS from P. intermedia ATCC 25611 was prepared by the standard hot phenol-water method. THP-1 cells were incubated in the medium supplemented with phorbol myristate acetate to induce differentiation into macrophage-like cells. The amount of TNF-alpha and interleukin-8 secreted into the culture medium was determined by enzyme-linked immunosorbent assay (ELISA). TNF-alpha and Ob-R mRNA expression levels were determined by semi-quantitative reverse transcription-polymerase chain reaction analysis.
RESULTS
Leptin enhanced P. intermedia LPS-induced TNF-alpha production in a dose-dependent manner. Leptin modulated P. intermedia LPS-induced TNF-alpha expression predominantly at the transcriptional level. Effect of leptin on P. intermedia LPS-induced TNF-alpha production was not mediated by the leptin receptor.
CONCLUSIONS
The ability of leptin to enhance P. intermedia LPS-induced TNF-alpha production may be important in the establishment of chronic lesion accompanied by osseous tissue destruction observed in inflammatory periodontal disease.

Keyword

Leptin; Lipopolysaccharide; Prevotella intermedia; Tumor necrosis factor-alpha

MeSH Terms

Body Weight
Cell Line
Enzyme-Linked Immunosorbent Assay
Humans
Inflammation
Interleukin-8
Leptin
Macrophages
Periodontal Diseases
Phorbols
Prevotella
Prevotella intermedia
Receptors, Leptin
RNA, Messenger
Tetradecanoylphorbol Acetate
Tumor Necrosis Factor-alpha
Interleukin-8
Leptin
Phorbols
RNA, Messenger
Receptors, Leptin
Tetradecanoylphorbol Acetate
Tumor Necrosis Factor-alpha

Figure

• 

  Figure 1 Effect of leptin on the release of tumor necrosis factor (TNF)-α and interleukin (IL)-8 by differentiated THP-1 cells. Cells were incubated with increasing concentrations of leptin (0.01-10 µg/mL) in the absence (A) or presence (B) of Prevotella intermedia lipopolysaccharide (LPS) (0.01 µg/mL). Supernatants were removed after 24 hours and assayed for TNF-α and IL-8. The results are means±SD of three independent experiments.

• 

  Figure 2 Effect of leptin on the expression of tumor necrosis factor (TNF)-α mRNA in differentiated THP-1 cells. Cells were incubated with increasing concentrations of leptin (0.01-10 µg/mL) in the absence (A) or presence (B) of Prevotella intermedia lipopolysaccharide (LPS) (0.01 µg/mL). See MATERIALS AND METHODS for further details. The polymerase chain reaction bands on a gel photograph in one of two separate experiments yielding similar results are shown.

• 

  Figure 3 Effect of leptin on the expression of Ob-Ra and Ob-Rb mRNA in differentiated THP-1 cells. Cells were incubated with increasing concentrations of leptin (0.01-10 µg/mL) in the absence (A) or presence (B) of Prevotella intermedia lipopolysaccharide (LPS) (0.01 µg/mL). See MATERIALS AND METHODS for further details. The polymerase chain reaction bands on a gel photograph in one of two separate experiments yielding similar results are shown.

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